芍药甘草汤修复PM2.5暴露系统性红斑狼疮小鼠的作用机制研究
Study on the mechanism of Shaoyao Gancao Decoction in repairing the damage of PM2.5-induced systemic lupus erythematosus mouse
金艺波 1蔡斯琦 1钱苏海 1范永升 1丁兴红1
作者信息
- 1. 浙江中医药大学基础医学院,杭州 310053
- 折叠
摘要
目的:建立PM2.5暴露系统性红斑狼疮(SLE)小鼠模型,研究芍药甘草汤对PM2.5致SLE小鼠病理损伤的治疗作用与机制.方法:40只C57BL/6雌性小鼠随机分为空白对照组、SLE组、SLE+PM2.5组(模型组)及芍药甘草汤组.模型组通过PM2.5全身暴露建立PM2.5暴露SLE小鼠模型.治疗结束后ELISA法检测小鼠血清抗ds-DNA抗体含量和肺泡灌洗液中活性氧(ROS)含量;HE染色观察肾脏和肺组织病理形态学改变;Western Blot检测相关蛋白NF-KBp65、p-NF-KBp65、TLR4、COX-2的表达水平;16SrRNA基因分析小鼠肠道微生物的变化.结果:与模型组比较,芍药甘草汤治疗后脾脏指数和血清抗ds-DNA抗体水平显著降低(P＜0.01);小鼠肾小球基底膜均匀、足突轻微融合;小鼠肺泡灌洗液中ROS含量降低(P＜0.05);HE染色切片发现小鼠肺组织肺泡炎症细胞渗出减少,NF-κBp65、p-NF-κB p65、TLR4、COX-2炎症相关蛋白水平显著降低(P＜0.05);16S rRNA基因分析结果显示,各组小鼠肠道微生物菌群具有多样性且组间结构存在差异性(P＜0.05),芍药甘草汤治疗后肠道菌群多样性提高,且保护性细菌丰度上调.结论:PM2.5可加重SLE发病,芍药甘草汤可改善PM2.5暴露后SLE小鼠的病理损伤,其作用机制可能是通过修复PM2.5诱导的炎症及肠道菌群紊乱对SLE发挥治疗作用.
Abstract
Objective:Established a systemic lupus erythematosus(SLE)mouse model exposured by PM2.5,and to study the therapeutic effect and mechanism of Shaoyao Gancao Decoction(SGT)on PM2.5 toxicity and pathological injury of(SLE)mice.Methods:A total of 40 C57BL/6 female mice were randomly divided into blank control group,SLE group,SLE+PM2.5 group(model group)and SGT group.Model group was exposed to PM2.5 to establish a mouse model of SLE.After treatment,the serum levels of anti-ds-DNA were measured by ELISA;the morphological changes of kidney and lung were observed by HE staining;the content of reactive oxygen species(ROS)in bronchoalveolar lavage fluid(BALF)was detected by ELISA.The related protein NF-κB p65,p-NF-κB p65,TLR4,COX-2 expression levels were detected by Western Blot.Collected mouse feces and analyzed the diversity and difference of intestinal microflora by 16S rRNA.Results:Compared with the model group,after treated with SGT,spleen index and the serum level of anti-ds-DNA were significantly decreased(P＜0.01);the glomerular basement membrane was uniform and the foot process was slightly fused in SGT group;the ROS content in BALF decreased significantly(P＜0.05);HE staining section showed that the inflammatory cell exudation was reduced,the inflammation related protein expression of NF-κB p65,p-NF-κB p65,TLR4,COX-2 were significantly decreased(P＜0.05).The results of 16S rRNA gene analysis showed that the intestinal microflora of mice in each group was diverse and the structure of each group was different(P＜0.05).The diversity of intestinal microflora was increased after treatment of SGT,and the abundance of protective bacteriawas up-regulated(P＜0.05).Conclusion:PM2.5 can exacerbate the incidence of SLE,SGT can improve the pathological damage of mice with SLE after PM2.5 exposure,the mechanism of action may be that it plays a therapeutic role in SLE by repairing PM2.5-induced inflammation and intestinal flora disorder.
关键词
系统性红斑狼疮/PM2.5/芍药甘草汤/炎症/肠道菌群Key words
Systemic lupus erythematosus/PM2.5/Shaoyao Gancao Decoction/Inflammation/Intestinal flora引用本文复制引用
基金项目
国家自然科学基金(81774179)
国家自然科学基金(81973778)
出版年
2024
NETL
NSTL
万方数据